Homework Answers
Answer:
In these type of questions, always remember: If a restriction enzyme has 1 cut size, in a circular plasmid, it will produce 'N' number of fragment, whereas in a linear DNA, it will produce 'N+1' number of fragments.
1. EcoR1 has 2 cut sites, and produces fragments of 2000bp and 5000bp
2. BamH1 has 1 cut site, it will linearize the DNA and produce a fragment of 5kb
3. Eae1 has 1 cut site, it will linearize the DNA and produce a fragment of 5kb
4. EcoR1+BamH1+Eae1 will have 4 cut sites in total, and produce fragments of sizes 1200bp, 800bp, 1000bp, and 2000bp
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The PCR was a success and your target region of 770 bp in length has been...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, dige...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
3. With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends...
3. With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends of the fragment that will be generated after digestion with Eael (indicate these in the two blocks below). What type of ends are generated by this enzyme (blunt/sticky, 5' or 3 overhang)? (4) GGCCR PCR amplicon RCCGG The PCR was a success and your target region of 770 bp in length has been amplified. You now plan to digest the DNA amplicon with the...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant plasmid containing the PCR fragment with the different restriction enzymes as listed. Recombinant plasmid Restriction enzyme No. of cuts Fragment sizes EcoRI BamHI Eael EcoRI + BamHI +Eae 1 -teamHI EcoRI 4000 EcoRI 6k6 1000 2000 es Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictch enzyme Egel, and clone the resulting longest fragment into...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictich enzyme Egel, and done the resulting longest fragment into Eael site of 5 KB plasmid. 4. You now run the four RE digests of the recombinant plasmid from question (4) on an agarose gel and stain it with ethidium bromide (EtBr). Indicate your expected profile on the gel below. Clearly indicate the...
Draw the plasmkd containing the cloned insert Indicate clearty where the insert will be ocated. I...
Please help. Every time I have posted this I have gotten a
different answer.
Draw the plasmkd containing the cloned insert Indicate clearty where the insert will be ocated. Include RE sites and distances. 300 EcoR3400 544kb 1000 2000 Cloned Insert The PCR was a success and your target region of 440 bp in length has been ampiied You igate a short linker containing an Apal restriction enzyme site onlo both ends of the PCR product, digest it with Apal...
Question 4 C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2:...
Question 4
C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2: Restriction enzyme digest analysis The PCR was a success and your target region of 440 bp in length has been amplified. You igate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below Bamll 300 EcoRI 3400 1000 2000 5...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb)....
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
Below is a set of experiments for cloning the human growth hormone (HGH) gene and then...
Below is a set of experiments for cloning the human growth hormone (HGH) gene and then expressing the HGH protein in E. coli starting from human pituitary glands and a tube of plasmid vector DNA. The plasmid expression vector contains the arabinose inducible expression system. Specify the correct order for carrying out these experiments, using the letter codes in front of each procedure. Use all of the steps in your answer, except for one step that should NOT be included....
please i need help with a, b, c this is the sequence 5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA...
please i need help with a, b, c
this is the sequence
5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA
AAGATCCTAACATTTTTGCGAG
TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC
AAATGGAAGTTTATATTTAAATA
GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA
ATAGTTGTGTAGATATAGGTCAT
GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA
ACAGTAATAATAACAATTTAAAACC
AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA
AACCAGAGTACAATAATAACAATT-3’
1. Polymerase chain reaction 5'- ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA ATAGTTGTGTAGATATAGGTCAT GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA ACAGTAATAATAACAATTTAAAACC AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA AACCAGAGTACAATAATAACAATT-3' a) One strand of a chromosomal DNA sequence is shown above. How would you amplify and isolate a DNA fragment defined by the sequence shown in red, using polymerase chain reaction. Design PCR primers (Forward and Reverse primers, each 20 nucleotides long, that...
The PCR was a success and your target region of 440 bp in length has been amplified. You ligate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below amHI 300 EcoRI 5 kb 3400 1000 EcoRI 2000 Apal Draw the plasmid containing the cloned insert. Indicate clearly where the insert will be located. Include RE...
3. With reference to your PCR amplicon, detail the terminal DNA sequences at the two ends of the fragment that will be generated after digestion with Eael (indicate these in the two blocks below). What type of ends are generated by this enzyme (blunt/sticky, 5' or 3 overhang)? (4) GGCCR PCR amplicon RCCGG The PCR was a success and your target region of 770 bp in length has been amplified. You now plan to digest the DNA amplicon with the...
UuOU Wuuur 3. Complete the table below to show your anticipated results, when digesting the recombinant plasmid containing the PCR fragment with the different restriction enzymes as listed. Recombinant plasmid Restriction enzyme No. of cuts Fragment sizes EcoRI BamHI Eael EcoRI + BamHI +Eae 1 -teamHI EcoRI 4000 EcoRI 6k6 1000 2000 es Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictch enzyme Egel, and clone the resulting longest fragment into...
- BamHI Ecort 4000 400 EcoRI 5kb 100 2000 I Target region of 770 bp in length has been amplified Plan to digest the DNA amplican with restrictich enzyme Egel, and done the resulting longest fragment into Eael site of 5 KB plasmid. 4. You now run the four RE digests of the recombinant plasmid from question (4) on an agarose gel and stain it with ethidium bromide (EtBr). Indicate your expected profile on the gel below. Clearly indicate the...
Please help. Every time I have posted this I have gotten a
different answer.
Draw the plasmkd containing the cloned insert Indicate clearty where the insert will be ocated. Include RE sites and distances. 300 EcoR3400 544kb 1000 2000 Cloned Insert The PCR was a success and your target region of 440 bp in length has been ampiied You igate a short linker containing an Apal restriction enzyme site onlo both ends of the PCR product, digest it with Apal...
Question 4
C. Cloning and restriction enzyme digest Video aid 1: Plasmid cloning Video aid 2: Restriction enzyme digest analysis The PCR was a success and your target region of 440 bp in length has been amplified. You igate a short linker containing an Apal restriction enzyme site onto both ends of the PCR product, digest it with Apal, and clone it into the Apal site of the 5 kb plasmid diagrammed below Bamll 300 EcoRI 3400 1000 2000 5...
Restriction Mapping Below is a restriction map for the plasmid PGEN101 (total length - 20 kb). Using this map as a guide, give the number of restriction fragments along with their associated lengths that would result from digesting PGEN101 with the restriction enzymes EcoRI, BamHII, anda combination of EcoRI + BamHI. BamHI BamHI BamHI / PGEN101 (20 kb) Mb EcoRI Digest Performed Size Emments Obtained EcoRI........ BamHI.. EcoRI + BamHI.... Two freshmen college students, interested in becoming gene jocks, performed...
III. Subclone the gene into plasmid, extract the plasmid DNA. 5. You know that your insert (gene of interest, GOI) is flanked by the EcoRI sites, which makes this restriction enzyme a perfect candidate to cut out your gene. You also know that the GOI has a unique BamH1 restriction site. After subcloning the PCR product into the plasmid, a purified DNA preparation of the plasmid is digested to completion with BamHI restriction endonuclease. In separate reactions, the same preparation...
please i need help with a, b, c
this is the sequence
5’ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA
AAGATCCTAACATTTTTGCGAG
TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC
AAATGGAAGTTTATATTTAAATA
GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA
ATAGTTGTGTAGATATAGGTCAT
GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA
ACAGTAATAATAACAATTTAAAACC
AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA
AACCAGAGTACAATAATAACAATT-3’
1. Polymerase chain reaction 5'- ATGTATTATTATTTTTTTGTTTTTTTTGCAATATATGCTAATGGATTGCTAAGAAATA AAGATCCTAACATTTTTGCGAG TAGCAATGATGAGATCATAGAAAATGATAAAAGTATGAATACCTTTGTTATGTCAAC AAATGGAAGTTTATATTTAAATA GTGATTTTAATTTAAATGAAGCATCCAACGAAAGCTTCTTAGAAAATTGCAATATCA ATAGTTGTGTAGATATAGGTCAT GAAAATGGCAACAAAATAAATAGTCAAGAAAATGAGCATGCTAAAAATAATAACA ACAGTAATAATAACAATTTAAAACC AGAATACAATAATAATAATAATAATTTAAAACCAGAATACAATAATAATAATTTAA AACCAGAGTACAATAATAACAATT-3' a) One strand of a chromosomal DNA sequence is shown above. How would you amplify and isolate a DNA fragment defined by the sequence shown in red, using polymerase chain reaction. Design PCR primers (Forward and Reverse primers, each 20 nucleotides long, that...
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